Yeast Mannoproteins Research Articles

Impact of the Chemical Composition and Structure of Yeast Mannoproteins from Pichia Strains on Wine Fruity Ester Release

Impact of the Chemical Composition and Structure of Yeast Mannoproteins from <i>Pichia</i> Strains on Wine Fruity Ester Release

Identification of the Yeast Mannoprotein Gene HZY1 as a Key Genetic Determinant for Yeast-Derived Haze in Beer

With the sustained popularity of hazy IPAs, brewers have explored multiple approaches to maximizing stable haze that will remain in suspension throughout the shelf life of the beer. Our recent investigations into yeast-dependent haze have uncovered specific brewing yeast strains that promote the formation of haze in heavily dry-hopped beer styles. These brewing strains have been termed “haze-positive” and furthermore, the timing of dry hop additions has been found to be another key factor in producing this stable haze. Classical genetics have identified YIL169C (herein referred to as HZY1) as both necessary and sufficient for the haze-positive phenotype in the yeast strain most widely used for Hazy IPAs. HZY1 encodes a candidate glycoprotein and our recent findings suggest it is localized to the cell wall through a GPI anchor. Surprisingly, using long-read sequencing data we uncovered extensive genetic variation in HZY1 across brewing strains. The haze-positive phenotype correlates with an expansion in the N-terminal serine-rich region. We propose that the Hzy1 glycoprotein is a critical component to yeast-dependent colloidal haze and the genetic variation in this locus contributes the range of haze phenotypes observed across industrial brewing strains.

Evaluation of the Addition of Yeast Mannoprotein to Oenococcus oeni Starter Cultures to Improve Wine Malolactic Fermentation

Malolactic fermentation (MLF) in wine is driven by the lactic acid bacterium Oenococcus oeni in most cases. Although this bacterium is resistant to wine stress conditions, it often faces difficulties completing MLF. Previous studies indicate that yeast mannoproteins may improve O. oeni growth and survival in wine. However, very little is known about this topic. This study evaluated the effect of the addition of mannoprotein extracts to culture media on O. oeni growth and its survival to stress conditions and MLF performance. Three commercial mannoprotein extracts were characterized in terms of polysaccharide and protein richness and were used for O. oeni culture media supplementation. The addition of mannoprotein extracts improved the survival of the two evaluated O. oeni strains, PSU-1 and VP41, after acid shock (pH 3.2) in comparison to that of the control. The transcriptional response of four genes involved in mannose metabolism was different depending on the strain, indicating the complexity of sugar metabolism in O. oeni. PSU-1 cells grown with two of the mannoprotein extracts performed faster MLF compared with the control condition, indicating that mannoprotein addition may improve the performance of O. oeni starter cultures, although this effect depends on the strain.

Interactions between Starmerella bacillaris and Saccharomyces cerevisiae during sequential fermentations influence the release of yeast mannoproteins and impact the protein stability of an unstable wine

Wine protein haze formation is a problem due to grape proteins aggregation during wine storage. The cell wall components of wine yeasts, particularly high molecular weight mannoproteins, have a protective effect against haze formation, although their involvement remains poorly understood. This study aimed at characterizing glycosylated proteins released by Starmerella bacillaris and Saccharomyces cerevisiae during single and sequential fermentations in a synthetic must, and testing their impact on wine protein stability. Mannoproteins-rich extracts from sequential fermentations showed an increase in the low MW polysaccharide fraction and, when added to an unstable wine, had a greater effect on protein stability than S. cerevisiae extracts. Shotgun proteomics approaches revealed that the identified cell wall proteins exclusively found in sequential fermentations were produced by both S. bacillaris (MKC7, ENG1) and S. cerevisiae (Bgl2p). Moreover, sequential fermentations significantly increased the expression of Scw4p and 1,3-beta-glucanosyltransferase (GAS5), produced by S. cerevisiae. Finally, some of the key proteins identified might play a positive role in increasing wine protein stability.

Obtaining yeast mannoproteins with antimicrobial properties

Obtaining yeast mannoproteins with antimicrobial properties

Evaluation of the Long-Lasting Flavour Perception after the Consumption of Wines Treated with Different Types of Oenological Additives Considering Individual 6-n-Propylthiouracil Taster Status.

Due to the limited scientific knowledge on the impact of commercial oenological additives on flavour perception, the aim of this work was to evaluate the effect of different types of oenological additives on the long-lasting flavour perception (flavour persistence) during wine tasting, also considering the effect of the individual PROP (6-n-propylthiouracil) taster status (PTS). To do so, white and red wines with two oenotannins (ellagitannin and gallotannin) and a commercial yeast mannoprotein were prepared. A control wine of each type was also made without additives. All the wines were spiked with a mixture of aromatic compounds responsible for the "fruity" and "woody" notes. Retronasal aroma and astringency were evaluated at the same time using time-intensity (TI) methodology and a trained panel (n = 40), including PROP non-tasters (NTs) and tasters (Ts). The results showed a significant effect of PTS on the long-lasting perception of astringency, being Ts who showed higher values than NTs for most TI parameters. However, PTS did not affect aroma persistence. In addition, the three oenological additives had an effect on astringency and retronasal aroma perception. They significantly increased the long-lasting perception of astringency compared to the control, while gallotannin also increased the persistence of the woody aroma.

Quality Characteristics of Reduced-Fat Emulsified Sausages Made with Yeast Mannoprotein Enzymatically Prepared with a β-1,6-glucanase

Mannoproteins, as yeast polysaccharides, have been utilized in food the industry as dietary fibers, emulsifying agents or fat replacers. Mannoprotein MP112, produced from yeast by enzymatic hydrolysis of myxobacterial β-1,6-glucanase GluM, exhibits excellent emulsifying properties in emulsion preparation. In this study, we aimed to examine the application of stable emulsion with the addition of mannoprotein MP112 (MP112 emulsion) to reduce the fat content of sausages. The addition of MP112 emulsion in emulsified sausages significantly reduced the fat content and increased the moisture and protein contents of emulsified sausages without the expense of their good sensory quality. Moreover, the textural properties of sausages were markedly improved with the higher hardness, chewiness and cohesiveness, especially in the 50–75% replacement ratio of MP112 emulsion. On the other hand, MP112 emulsion replacement of animal fat markedly improved the nutritional composition of emulsified sausages; they displayed a higher PUFA/SFA ratio and lower n-6/n-3 ratio due to their saturated fatty acids being replaced by poly-unsaturated fatty acids. Meanwhile, the oxidative stability of sausages was improved linearly, corresponding to the increased replacement ratio of MP112 emulsion. Our results show that mannoprotein-based emulsions could be used as potential fat alternatives in developing reduced-fat meat products.

Yeast mannoproteins: Organoleptic modulating functions, mechanisms, and product development trends in winemaking

AbstractMannoproteins (MPs) originate from yeast cells and can play important roles in maintaining wine stability and modulating wine organoleptic properties. Due to their natural presence in wines, MP products are allowed to be used as enological additives in wine industry. To guarantee an appropriate application of these products during winemaking, it is necessary for both researchers and winemakers to understand the mechanisms of MPs as multifunctional organoleptic modulators. This review has introduced the current state of knowledge of the researches focus on the application of different MP products in red wines and also the interactions between MPs and wine sensory active components (polyphenols, aroma compounds) in model wine solutions. We have summarized the possible mechanisms of MPs that affect the color, astringency, and aroma of red wines and the relationship between the physicochemical characteristics and functionalities of MPs. Besides, development trends of MP products are also discussed. It seems that yeast biomass, especially those from non‐Saccharomyces yeasts show good potential in producing large quantities of MPs, an appropriate combination of yeast cultivation and preservation methods with extraction and purification techniques may obtain MPs of ideal production, purity, and functionality in an industrial scale, which should be put more efforts by both scientists and manufacturers in the future.

Brewer's Spent Yeast Cell Wall Polysaccharides as Vegan and Clean Label Additives for Mayonnaise Formulation.

Brewer's spent yeast (BSY) mannoproteins have been reported to possess thickening and emulsifying properties. The commercial interest in yeast mannoproteins might be boosted considering the consolidation of their properties supported by structure/function relationships. This work aimed to attest the use of extracted BSY mannoproteins as a clean label and vegan source of ingredients for the replacement of food additives and protein from animal sources. To achieve this, structure/function relationships were performed by isolating polysaccharides with distinct structural features from BSY, either by using alkaline extraction (mild treatment) or subcritical water extraction (SWE) using microwave technology (hard treatment), and assessment of their emulsifying properties. Alkaline extractions solubilized mostly highly branched mannoproteins (N-linked type; 75%) and glycogen (25%), while SWE solubilized mannoproteins with short mannan chains (O-linked type; 55%) and (1→4)- and (β1→3)-linked glucans, 33 and 12%, respectively. Extracts with high protein content yielded the most stable emulsions obtained by hand shaking, while the extracts composed of short chain mannans and β-glucans yielded the best emulsions by using ultraturrax stirring. β-Glucans and O-linked mannoproteins were found to contribute to emulsion stability by preventing Ostwald ripening. When applied in mayonnaise model emulsions, BSY extracts presented higher stability and yet similar texture properties as the reference emulsifiers. When used in a mayonnaise formulation, the BSY extracts were also able to replace egg yolk and modified starch (E1422) at 1/3 of their concentration. This shows that BSY alkali soluble mannoproteins and subcritical water extracted β-glucans can be used as replacers of animal protein and additives in sauces.

Cell-surface protein YwfG of Lactococcus lactis binds to α-1,2-linked mannose.

Lactococcus lactis strains are used as starter cultures in the production of fermented dairy and vegetable foods, but the species also occurs in other niches such as plant material. Lactococcus lactis subsp. lactis G50 (G50) is a plant-derived strain and potential candidate probiotics. Western blotting of cell-wall proteins using antibodies generated against whole G50 cells detected a 120-kDa protein. MALDI-TOF MS analysis identified it as YwfG, a Leu-Pro-any-Thr-Gly cell-wall-anchor-domain-containing protein. Based on a predicted domain structure, a recombinant YwfG variant covering the N-terminal half (aa 28-511) of YwfG (YwfG28-511) was crystallized and the crystal structure was determined. The structure consisted of an L-type lectin domain, a mucin-binding protein domain, and a mucus-binding protein repeat. Recombinant YwfG variants containing combinations of these domains (YwfG28-270, YwfG28-336, YwfG28-511, MubR4) were prepared and their interactions with monosaccharides were examined by isothermal titration calorimetry; the only interaction observed was between YwfG28-270, which contained the L-type lectin domain, and d-mannose. Among four mannobioses, α-1,2-mannobiose had the highest affinity for YwfG28-270 (dissociation constant = 34 μM). YwfG28-270 also interacted with yeast mannoproteins and yeast mannan. Soaking of the crystals of YwfG28-511 with mannose or α-1,2-mannobiose revealed that both sugars bound to the L-type lectin domain in a similar manner, although the presence of the mucin-binding protein domain and the mucus-binding protein repeat within the recombinant protein inhibited the interaction between the L-type lectin domain and mannose residues. Three of the YwfG variants (except MubR4) induced aggregation of yeast cells. Strain G50 also induced aggregation of yeast cells, which was abolished by deletion of ywfG from G50, suggesting that surface YwfG contributes to the interaction with yeast cells. These findings provide new structural and functional insights into the interaction between L. lactis and its ecological niche via binding of the cell-surface protein YwfG with mannose.

Study on the mechanism of interaction between mulberry anthocyanins and yeast mannoprotein

To solve the problem of mulberry juice colour instability caused by ultraviolet radiation, we proposed the addition of mannoprotein (MP) to improve the stabilization of mulberry juice colour. First, the purified mulberry anthocyanins extracted (MAE) were identified by UFLC–MS, and their purity was determined by liquid-phase peak area normalization. The interaction mechanism between MP and MAE at different pH values was systematically investigated by Fourier transform infrared (FTIR) spectroscopy, circular dichroism (CD) spectroscopy, fluorescence spectroscopy, and molecular docking. After conjugation with MAE, the secondary structure of MP changed slightly. The fluorescence quenching of MP by MAE was dose dependent. Thermodynamic analysis showed that the main interaction forces at pH 3.2 were hydrogen bonding and van der Waals forces, while the main force at pH 7.4 was hydrophobic forces. The molecular docking results verified the good binding properties of MP with cyanidin-3-O-glucoside (C3G) and cyanidin-3-O-rutinoside (C3R).

Antimicrobial and prebiotic activity of mannoproteins isolated from conventional and nonconventional yeast species—the study on selected microorganisms

Yeast mannoproteins are proposed as a paraprobiotics with antimicrobial and prebiotic properties. They can be used as biopreservatives in food and in diseases therapies. The knowledge about the specificity and/or capability of their influence on the growth of different microorganism is limited. The study determined the effect of mannoprotein preparations of Saccharomyces cerevisiae (S. cerevisiae) ATCC 7090 and nonconventional yeast origin [Metschnikowia reukaufii (M. reukaufii) WLP 4650 and Wickerhamomyces anomalus (W. anomalus) CCY 38-1-13] on the growth of selected bacteria of the genera: Lactobacilllus, Limosilatobacillus, Limosilatobacillus, Bifidobacterium, Staphylococcus, Enterococcus, Pseudomonas, Escherichia, Proteus and Salmonella. The degree of stimulation or growth inhibition of tested bacteria depended on the type and dose of the mannoprotein and the bacterial strain. The addition of the tested preparations in the entire range of applied concentrations had a positive effect especially on the growth of Lactobacillus arabinosus ATCC 8014 and Bifidobacterium animalis subsp. lactis B12. Mannoproteins isolated from S. cerevisiae limited the growth of the Escherichia coli (E. coli) ATCC 25922, Pseudomonas aureoginosa (P. aureoginosa) ATCC 27853, Proteus mirabilis ATCC 35659 and Salmonella Enteritidis ATCC 13076 to the greatest extent, while preparations of M. reukaufii and W. anomalus origin most effectively limited the growth of Staphylococcus aureus strains, E. coli and P. aureoginosa. The growth of Enterococcus faecalis was stimulated by the presence of all studied preparations in most of the concentrations used. Further research will determine how the purification process of studied mannoproteins or oligosaccharide fractions, its structure and composition influence on the growth of selected bacteria and what is the mechanism of its activity.

Yeast mannoproteins are expected to be a novel potential functional food for attenuation of obesity and modulation of gut microbiota.

The yeast mannoproteins (MPs), a major component of yeast cell walls with large exploration potentiality, have been attracting increasing attention due to their beneficial effects. However, the information about the anti-obesogenic activity of MPs is still limited. Thus, the effects of MPs on the high-fat diet (HFD)-induced obesity and dysbiosis of gut microbiota were investigated in this work. The results showed that MPs could significantly attenuate the HFD-induced higher body weight, fat accumulation, liver steatosis, and damage. Simultaneously, the inflammation in HFD-induced mice was also ameliorated by MPs. The pyrosequencing analysis showed that intervention by MPs could lead to an obvious change in the structure of gut microbiota. Furthermore, the prevention of obesity by MPs is highly linked to the promotion of Parabacteroides distasonis (increased from 0.39 ± 0.12% to 2.10 ± 0.20%) and inhibition of Lactobacillus (decreased from 19.99 ± 3.94% to 2.68 ± 0.77%). Moreover, the increased level of acetate (increased from 3.28 ± 0.22 mmol/g to 7.84 ± 0.96 mmol/g) and activation of G protein-coupled receptors (GPRs) by MPs may also contribute to the prevention of obesity. Thus, our preliminary findings revealed that MPs from yeast could be explored as potential prebiotics to modulate the gut microbiota and prevent HFD-induced obesity.

Structural characterization and emulsifier property of yeast mannoprotein enzymatically prepared with a β-1,6-glucanase

Yeast cell wall mannoprotein with diverse structures and functions are effective emulsifiers due to the amphiphilic structure comprising a protein backbone and mannan chains. In this study, a β-1,6-glucanase-based approach to produce mannoproteins from baker's yeast was established. Batch production of mannoprotein MP112 was realized with 11.1% yield, 90.7% purity, and a mannan to protein ratio of 14.5. The mannan-protein conjugate structure was confirmed by composition analysis, FT-IR and NMR spectrometry, as well as SEM observation, and the assembled mesoporous structure with filamentous fibers was deduced to determine the favorable emulsifying properties from rheological analysis. Application of mannoprotein-based emulsion in the preparation of porcine myofibrillar protein gels showed that MP112 improves the properties of myofibrillar protein composite gels by the formation of a fine-stranded gel network with enhanced protein-polysaccharide-water interactions, which reduce damage to the muscle tissue and increase product quality. This study is expected to broaden the applications of mannoproteins as value-added ingredients.

Role of Yeast Mannoproteins in the Interaction between Salivary Proteins and Flavan-3-ols in a Cell-Based Model of the Oral Epithelium.

Astringency is a highly complex sensation which involves multiple mechanisms occurring simultaneously, such as the interaction between flavan-3-ols and salivary proteins (SP). Moreover, astringency development can be affected by the presence of polysaccharides such as mannoproteins (MP). The aim of this work was to evaluate the molecular mechanisms whereby MP could modulate the astringency elicited by tannins, using a cell-based model of the oral epithelium (TR146 cells), and the effect of salivary proteins on these interactions. The binding of flavan-3-ols to oral cells was evaluated by DMACA assay, while the content of unbound flavan-3-ols after the interactions was assessed by means of HPLC-DAD-MS. Results obtained confirm the existence of cell–tannin interactions, that can be partially inhibited by the presence of SP and/or MP. The most significant decrease was obtained in the system containing MPF (38.16%). Both mannoproteins assayed seem to have modulating effect on flavan-3-ol–SP interactions, acting by two different mechanisms: MPF would lead to the formation of SP/MPF/flavan-3-ols ternary soluble aggregates, while MPL seems to prevent flavan-3-ol–saliva interaction by a competitive mechanism, i.e., MPL would reduce cell–tannin interactions, similar to SP. This study suggests that mannoproteins with different compositional characteristics could exhibit preferential interaction with distinct flavan-3-ol families.

Selection of Calabrian strains of Saccharomyces sensu stricto for red wines

AbstractPhenolic compounds provide important quality attributes to red wines interacting with the organoleptic impact of wines. Yeast mannoproteins can interact with grape phenolic compounds, responsible for colour and antioxidant activity of wines. The aim of this work was to perform oenological characterisation and specific selection of Calabrian strains ofSaccharomyces sensu stricto. Among the considered traits, the aptitude of the yeast to preserve grape pigments and colour intensity was included. Among the best six yeast strains – Sc2731, Sc2742, Sc2756, Sc2773, Sc2774, and Sc2823 – strain Sc2742 exhibits the highest Folin–Ciocalteu index and strain Sc2774 the highest colour intensity. These two selected yeasts may be used as starter for the production of red wines in order to preserve grape pigments and colour intensity.

Extraction of mannoprotein from Saccharomyces cerevisiae and analysis of its chemical composition and molecular structure

The purpose of this study was to screen the method with the highest yield of mannoprotein (MP) without damaging their functional structure. The recovery rates of the extracts, protein, and mannose were determined along with the mannose/protein ratio, molecular weight and distribution, monosaccharide and amino acid compositions, and secondary structures of the three MP extracts to compare the extraction methods. The MP extracts recovery rate prepared by the thermal method was significantly higher (35.89%) than those obtained using fermentation (31.66%) and SDS treatment (19.77%). Three protein bands with similar molecular weights of 59, 47, and 34 kDa were detected in the MPs obtained via the different extraction methods. The thermally extracted MP has a broader molecular weight distribution. After purification, the proportion of mannose in the polysaccharide parts of the MPs increased from 6–7 to 90.4–91.3%. The essential amino acid content of the hot-extracted MP (170.07 mg/g) was the highest. The thermally extracted MP had similar secondary structural characteristics to that isolated at room temperature, and had a higher protein characteristic peak intensity. In general, the heating method to extract yeast mannoprotein is time-saving and efficient.

Use of Yeast Mannoproteins by Oenococcus oeni during Malolactic Fermentation under Different Oenological Conditions.

Oenococcus oeni is the main agent of malolactic fermentation in wine. This fermentation takes place after alcoholic fermentation, in a low nutrient medium where ethanol and other inhibitor compounds are present. In addition, some yeast-derived compounds such as mannoproteins can be stimulatory for O. oeni. The mannoprotein concentration in wine depends on the fermenting yeasts, and non-Saccharomyces in particular can increase it. As a result of the hydrolytic activity of O. oeni, these macromolecules can be degraded, and the released mannose can be taken up and used as an energy source by the bacterium. Here we look at mannoprotein consumption and the expression of four O. oeni genes related to mannose uptake (manA, manB, ptsI, and ptsH) in a wine-like medium supplemented with mannoproteins and in natural wines fermented with different yeasts. We observe a general gene upregulation in response to wine-like conditions and different consumption patterns in the studied media. O. oeni was able to consume mannoproteins in all the wines. This consumption was notably higher in natural wines, especially in T. delbrueckii and S. cerevisiae 3D wines, which presented the highest mannoprotein levels. Regardless of the general upregulation, it seems that mannoprotein degradation is more closely related to the fermenting medium.

Effect of different yeast mannoproteins on the interaction between wine flavanols and salivary proteins

Unbalanced wine astringency, caused by a gap between phenolic and technological grape maturities, is one of the consequences of the global climate change in the vitiviniculture. To resolve it, potential strategies are being currently used, like the addition of commercial yeast mannoproteins (MPs) to wines. In this work, the main interactions responsible for the wine astringent sensation, namely, interactions between human salivary proteins and wine flavanols have been studied by Dynamic Light Scattering (DLS) and liquid chromatography coupled to DAD and MS detectors (HPLC–DAD–MS), in presence or absence of two MPs with different saccharide/protein ratio. The results indicate that there are differences on the substrate specificity for each mannoprotein and that its action mechanism could change not only depending on the mannoprotein composition but also on the flavanol structure. MPs with elevated carbohydrate content could act thought the stabilization of soluble aggregates with human salivary proteins and flavanols, mainly non-galloylated flavanol oligomers, whereas MPs with higher protein percentage mostly could precipitate flavanols (mainly non-galloylated ones with low degree of polymerization) which partially prevents the formation of insoluble flavanol–salivary protein aggregates.

Influence of fermentation conditions on the secretion of seripauperin 5 (PAU5) by industrial sparkling wine strains of Saccharomyces cerevisiae

The yeast mannoprotein seripauperin 5 (PAU5) from Saccharomyces cerevisiae is a negative gushing biomarker in sparkling wine with a direct gushing-reducing effect. The knowledge about the influence of the yeast strain and the fermentation conditions on the PAU5 content in the final product could reduce the gushing potential of sparkling wines and avoid economic losses for sparkling wine producers. The potential of 30 different commercially used (sparkling) wine S. cerevisiae strains to produce PAU5 was analyzed by RP-HPLC. The experiments revealed great differences between strains and identified three high-PAU5-producing yeast strains. The influence of different fermentation conditions was analyzed in two selected strains. Cultivation conditions of elevated temperature and lower inoculation density, as well as cultivation under diffuse daylight significantly enhanced PAU5 production by the tested S. cerevisiae strains as compared to standard conditions. However, cultivation as agitated cultures and co-cultivation with Metchnikowia pulcherrima or Torulaspora delbrueckii led to a reduction of PAU5 production as compared to standard conditions.